摘要
目的:制备睫状神经营养因子(ciliary neurotrophic factor, C N T F)单克隆抗体。方法:将 C N T F免疫的 B A L B/c 小鼠脾细胞和 S P2/0 骨髓瘤细胞融合,经 3 次克隆化培养,获得分泌 C N T F单克隆抗体的杂交瘤细胞株;然后进行体内诱生获取含大量 C N T F单克隆抗体的腹水和血清;再进行 C N T F单克隆抗体鉴定和效价测定。结果:获得5 株分泌 C N T F单克隆抗体的杂交瘤细胞株,抗体亚型为 Ig G2a;腹水和血清中该抗体效价分别为 1∶107 ~1∶108 和 1∶106;该抗体能特异性地显示 S D大鼠脑内表达 C N T F的神经元,其中1 株单克隆抗体能抑制 C N T F促进体外培养新生 S D 大鼠骨骼肌细胞增殖的功能。结论:制备成功高效且特异的 C N T F单克隆抗体,为 C N T F结构、功能和机制的研究提供了有效工具。
Objective: To prepare the monoclonal antibodies against ciliary neurotrophic factor(CNTF). Methods: Cell lines of hybrids secreting monoclonal antibodies against CNTF were established by fusing SP2/0 myeloma cells with spleen cells of BALB/c mice immunized with CNTF and cloning 3 times; asitic fluid and serum containing abundant monoclonal antibodies against CNTF were obtained by in vivo induction; then identification of the monoclonal antibodies against CNTF and detection of antibody titer were undergone. Results: Five cell lines of hybrids secreting monoclonal antibodies against CNTF were established. Subtypes of these antibodies were all IgG 2a . Antibody titers in asitic fluid and serum were 1∶10 7~1∶10 8 and 1∶10 6, respectively. These monoclonal antibodies showed specificity in demonstrating neurons secreting CNTF in CNS, and one kind of them inhibited the enhancing effect of CNTF on the proliferation of neonal SD rats skeletal muscle cells in culture. Conclusion: The monoclonal antibodies against CNTF with high activity and specificity have been established successfully, and will be a useful tool in the studies of CNTF's structure,function and mechanism
出处
《第二军医大学学报》
CAS
CSCD
北大核心
1999年第8期560-562,共3页
Academic Journal of Second Military Medical University